James Wakefield

My research interest has always been that of mitosis and cell division, stimulated by the fundamental beauty of the process as viewed using a fluorescence microscope, and its key role in diseases such as cancer. When I was an "A" level student I had an inspirational Biology teacher, Alan Wright, who told me that one day, when I grew up, I should go and find out how microtubules organised themselves during mitosis. I'm still working on an explanation - but then I'm not yet really grown up……..

Jack Chen - Post-doc

There are three major areas of microtubule nucleation in eukaryotic cells. They are the centrosomes, chromosomes, and pre-existing microtubules. The Augmin complex, consisting of 8 different proteins, is responsible for regulating microtubule generation around pre-existing microtubules. The current predominant model is that Augmin recruits γ-Tubulin Ring Complex to the microtubules which then nucleates new microtubules.

Using an in vitro approach, I am purifying GFP-tagged Augmin, to see if and where it binds to microtubules. I am also seeing if Augmin can nucleate new microtubules. Using an add-back approach, I will determine if human Augmin and drosophila Augmin are functionally conserved. Using an in vivo approach I will examine the role of Augmin in various drosophila tissues, including embryos, oocytes, and neuroblasts, to determine the role of Augmin in generating microtubules not only in mitosis, but also in specialized cell division, and during cell differentiation.

Before joining the Wakefield lab, I did my masters with Dr. Graham Dellaire at Dalhousie University, Halifax, Canada. My research project was on the regulation of promyelocytic leukemia protein phosphorylation, and its interaction with other proteins, in the DNA damage response pathway.

My special skills include being quasi-ambidextrous and able to fence with either hands, and being a pundit at cracking eggcellent jokes.

Stacey Scott - MRes Student

Cell division is a fundamental process driven by the formation of a microtubule based mitotic spindle. Defects within this structure can lead to chromosome instability and problems with cell polarisation. Understanding how microtubules are formed and how they are regulated is important in understanding a number of different diseases, one such example being cancer. Nucleation, length and dynamics of microtubules are determined by microtubule associated proteins (MAP’s). Originally identified in mammalian brain tissues MAPs can be defined as a protein that forms associations with microtubules, their cargos or motors. The main aims of my project are to apply quantitative proteomics and biochemistry in order to characterise a number of MAPs in the early Drosophila embryo. Buy carrying out immunoprecipitation and mass spectrometry, in combination with fluorescence microscopy of Drosophila melanogaster embryos expressing a GFP fusion of our protein of interest; we are aiming to assess both localisation and function of selected mitotic MAPs. Before starting my Masters by research in the Wakefield Lab, in 2013 I graduated from the University of Exeter with an upper second class degree with honours in Human Biosciences; my third year project focusing on the role of dynein in the motility of the early endosome in the pathogenic fungus U.maydis with Professor G. Steinberg. In October 2015 I will be moving to Cambridge, to take up a PhD with Paolo D'Avino. When not in the lab I can usually be found ballroom dancing, running or being overly enthusiastic about penguins.

Lizzy Anderson - MRes Student

During mitosis various proteins interact with microtubules as MAPs to aid the process. As an undergraduate I researched the possibility of a Drosophila protein called stem loop-binding protein (Slbp) as being a MAP during mitosis. Slbp has previously been characterised to have a role in histone pre-mRNA processing, however, I was able to visualise a GFP-tagged version of Slbp in the region of the microtubules during mitosis. Further to this, mass spectrometry analysis identified a large number of specific interacting proteins, with various functions, suggesting that Slbp may be playing a larger role.

As an MRes student I am now investigating further the role that Slbp plays in mitosis through both in vitro and in vivo methods. This work will be conducted using the model organism Drosophila melanogaster. Due to the known deficiency of Slbp in patients with Wolf-Hirschorn syndrome, I hope to go on to work with human cell lines in the future.

When I am not in the lab I am a keen baker and runner (to counter-act the baking!), and I enjoy music and doing open-mic nights around Exeter.

James Marks - MRes / PhD Student

The mitotic spindle, a fundamental cellular structure composed of microtubules (MTs) and associated proteins (MAPs), ensures correct alignment and segregation chromosomes during mitosis. It is generated through a series of steps that can be defined as MT nucleation, MT coalescence and spindle bipolarisation, involving a set of conserved microtubule motors which dynamically organise the spindle. As part of a Masters by research I will look to identify the roles of an array of GFP tagged motor proteins known to function during Drosophila spindle formation. I will observe their localisations during the cell cycle, immunopurify them for mass spectrometry to identify interacting proteins, and undertake in vitro studies with the purified soluble protein complexes to determine their effects on MT dynamics, nucleation and bundling.

As of October 2015, I will be beginning a multi-disciplinary BBSRC-funded PhD, continuing my MRes work, with James Wakefield and Jeremy Metz.

Outside the lab I am a keen classical guitar player, and dabble in jazz when given the chance, and enjoy skiing, brewing and kayaking.

Past Members

Daniel Hayward - PhD Student

Dan was a PhD student in the lab between 2010-2013.

His research focused on the multiple apparatus that contribute to the building of the mitotic spindle, and the pathways and molecules that underpin these mechanisms. His model system is the single celled, multi-nuclei Drosophila early syncytial embryo, enabling him to use high resolution confocal microscopy to obtain 4-dimensional images of mitotic spindles forming in vivo. This system also allows the use of biochemistry, proteomics, genetics and interfering antibody injection.

His work was recently published in Developmental Cell (Hayward et al., 2014) and in Communicative and Integrative Biology (Hayward and Wakefield, 2014).

Dan is currently a post-doc with Ulli Gruneberg at the Dunn School of Pathology, University of Oxford, where he is looking at the role of post-translational modifications on protein function during spindle formation.

Pete Jones - Post-Doc

Pete was a post-doc in the lab between 2010 and 2013.

The chromosomal passenger complex (CPC) is a key regulator of cell division and has crucial roles throughout the cell cycle. This complex consists of the Aurora B kinase and three other subunits – Incenp, Borealin and Survivin – which are essential for its highly dynamic localisation (see movie) and activity. The CPC regulates various stages of cell division – spindle assembly, chromosome alignment, correction of microtubule-kinetochore attachment errors, central spindle and ultimately cytokinesis. Pete used proteomic approaches to attempt to identify novel Aurora B substrates and interactors of the complex.

Pete is now working in the Department of Cardiovascular Sciences at the University of Leicester

Faisal Khan - PhD Student

Faisal was a PhD student in the lab, jointly supervised by Charlotte Deane's Bioinformatics lab in Oxford, between 2009-2013.

Protein-protein interaction (PPI) networks are becoming increasingly popular guides to help biologists identify and target uncharacterised proteins that might possess a putative function in different biological processes. In my inter-disciplinary project, we created an extended PPI network, or interactome, for mitotic microtubule-associated proteins (MAPs) in Drosophila. Each protein, or node, in this network was annotated with different experimental and bioinformatics data, and then using a statistical model, we scored each protein based on the predicted likelihood of their involvement in mitosis. By combining and using multiple types of available biological data, our interactome not only predicts the likelihood of a mitotic role for our putative MAPs but also unravels the inter-relationship of different proteins and their functions. This gives us an integrated view of the process and guides our in vivo study of novel mitotic proteins. I am now chasing some very interesting candidate proteins (back in the wetlab) using an array of methods from biochemistry, cell biology and genetics.

Faisal is now the Director of the Institute of Integrative Biosciences at CECOS University in Pakistan.